Neurodegenerative diseases have become enormous financial and societal burdens as human life expectancy has increased. Misregulation of autophagy has been implicated in neurodegenerative disorders, including Alzheimer's disease, Parkinson's disease and Huntington's disease. While many studies have characterized autophagy and its role in cellular homeostasis in yeast and mammalian cell culture, much less in known about autophagy in neurons, especially in the context of intact, living animals. Autophagosome biogenesis decreases with age in mammalian primary neurons; the kinetics of autophagosome assembly factors also become disrupted in primary neurons from aged mice (Stavoe et al., under peer review). Understanding how all stages of autophagy change with age and how autophagy is regulated during aging will be important for developing treatments for age-related neurodegenerative diseases. Furthermore, uncovering how aging modulates autophagy in living animals, with intact nervous systems, will take into account the complex, dynamic natural environment in which neurodegenerative diseases develop. The identification of age-related alterations in the autophagy pathway and how autophagy is modulated with age will be conducted under the guidance of Dr. Erika Holzbaur (mentor, University of Pennsylvania), a world expert in neuronal autophagy and trafficking. The PI will collaborate with world experts in super-resolution microscopy (Drs. Melike Lakadamyali and Andrea Stout, UPenn) and viral gene transfer (Dr. John Wolfe, UPenn) to learn new state-of-the-art approaches to incorporate into the project. The PI has assembled an excellent research advisory committee who will provide the training, support, and mentoring to facilitate the proposed studies and growth of the PI. The PI will be based at the University of Pennsylvania, under the guidance of Dr. Erika Holzbaur during the entire period of the mentored award. In addition, the PI has access to state-of-the-art facilities and equipment, as well as excellent resources for career development at the University of Pennsylvania. The proposed research and development plan will enable the PI to characterize the age-related alterations to the autophagy pathway and determine how autophagy is regulated in both primary mammalian neurons and C. elegans neurons in vivo. This initial research will allow the PI to then modulate neuronal autophagy in vivo in both murine and C. elegans models of aging and neurodegeneration, thus creating a niche for the PI as an independent faculty member. These studies will determine potential molecular targets for improving the healthspan of the nervous system in the context of aging and neurodegeneration. The proposed studies are part of the PI's long-term goal to investigate the evolutionarily conserved molecular mechanisms of neuronal aging and neurodegeneration, with a particular focus on neuronal autophagy, and to apply these discoveries to ameliorating neurodegenerative diseases.